Electroporation

yoginee budhkar via methods%40net.bio.net (by eenigoy from gmail.com)
Wed Jan 28 01:15:16 EST 2009


Dear All,

I want to use biorad gene pulser for transforming *E. coli* with my ligated
mix.

It is mentioned in the bio rad manual that we must incubate the ligated mix
at 65deg C for 15 min to inactivate the enzyme and then proceed with its
dilution or etOH precipitation to get rid of extra salts that may decrease
the resistance and cause arcing.

I did go for the boiling step, but I'm afraid I'll lose the DNA by
precipitation since already only about 80 to 100 ng, insert + vector DNA is
present in my ligated mix. Cannot afford to lose any of it during
precipitation

What could be done to get rid of the salt content that hampers the
electroporation?

-- 
--
Yoginee Budhkar
Junior Research Fellow
Food Engineering and Technology Department
Institute of Chemical Technology
Matunga, Mumbai 400019
Contact: +91 9223355677


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